Multiplatform Metabolomics to Understand the Imidacloprid-Induced Toxicity in Drosophila.

Neonicotinoids, the class of insecticides used for crop protection, are subjected to vigilance due to their pernicious impacts. Imidacloprid (IMD) is one of the most representative insecticides of the neonicotinoid family, which has shown unfriendly consequences for non-target species. Metabolomics, a multidisciplinary approach, is being used in toxicological research to understand the metabolic responses to toxicant exposure by utilizing modern analytical techniques. Yet, no solitary analytical technique can cover the broad metabolite spectrum, but a multi-technique metabolomics platform can aid in analyzing the majority of the metabolites. In the present study, an effort has been made to identify the differential metabolites in Drosophila after exposure to IMD at 2.5 and 25 ng/mL using liquid chromatography-high-resolution mass spectrometry (LC-HRMS), gas chromatography-MS (GC-MS), and NMR-based untargeted metabolomics. Multivariate pattern recognition analysis helped in identifying/recognizing 19 (LC-HRMS), 7 (GC-MS), and 13 (NMR) differential metabolites mainly belonging to the category of amino acids, sugars, fatty acids, and organic acids. The pathway analysis of differential metabolites predominantly showed impact on aminoacyl-tRNA biosynthesis, amino acid metabolism, and glycerophospholipid metabolism. Among these, arginine and proline metabolism was observed to be the common metabolic pathway perturbed in Drosophila due to IMD exposure. The multiplatform metabolomics based on LC-HRMS, GC-MS, and NMR analysis with an advanced level of statistical analysis can provide insights into potential perturbations in the metabolome of IMD-exposed Drosophila.

Phytometabolomic analysis of boiled rhizome of Nymphaea nouchali (Burm. f.) using UPLC-Q-TOF-MSE, LC-QqQ-MS & GC-MS and evaluation of antihyperglycemic and antioxidant activities.

Phytometabolomic analysis of Nymphaea nouchali (Burm. F.) boiled rhizome was carried out utilizing UPLC-Q-TOF-MSE, LC-QqQ-MS and GC-MS techniques and evaluated for antihyperglycemic and antioxidative stress potentials. Metabolomic analysis revealed presence of multiple antidiabetic and antioxidant compounds. Boiled rhizome powder exhibited potent antihyperglycemic activity against sugar-induced postprandial hyperglycemia in rats plausibly due to the presence of intestinal α-glucosidase inhibitory and augmenting cellular glucose uptake activities. It also prevented hyperglycemia-induced hemoglobin and insulin glycation. Rhizome displayed potent reducing power, effectively scavenged various reactive oxygen species. It displayed antioxidative stress potential in assuaging H2O2 induced erythrocyte hemolysis and antioxidant activity by inhibiting membrane lipid peroxidation. Boiled rhizome was also found to preserve the loss of cellular antioxidants under H2O2 induced oxidative stress and disturbances caused to mitochondrial membrane potential. This is the first research reporting boiled N. nouchali rhizome as an ideal food material to manage the cause of hyperglycemia and resultant oxidative stress.

Bombax ceiba (Linn.) calyxes ameliorate methylglyoxal-induced oxidative stress via modulation of RAGE expression: identification of active phytometabolites by GC-MS analysis.

Non-enzymatic reactions between proteins and methylglyoxal (MG) result in the formation of advanced glycation end products (AGEs). These AGEs play a vital role in the development of diabetic complications by stimulating oxidative stress and acting upon their receptor RAGE (Receptor for Advanced Glycation End products). This study examined the effect of aqueous methanol extract of Bombax ceiba L. calyxes (BCCE) on MG induced protein glycation and oxidative stress, followed by the identification of phytometabolites present in the calyxes using gas chromatography-mass spectrometry (GC-MS). The study revealed that priming of bovine serum albumin protein with the BCCE inhibited MG induced AGE formation in vitro and restrained AGE-induced RAGE up-regulation in HEK-293 cells. The BCCE significantly (p < 0.001) reduced the MG induced increase in reactive oxygen species (ROS), NADPH oxidase (NOX), and mitochondrial dysfunction. Improvements in the levels of antioxidant enzymes such as Mn and Cu/Zn-superoxide dismutase and glutathione reductase were also observed in HEK-293 cells. Furthermore, the decrease in primary cellular defense against AGEs, the glyoxalase 1 (Glo-1) activity, due to MG treatment was restored in BCCE treated cells. GC-MS analysis revealed the presence of antioxidant and antiglycation compounds such as myo-ionisitol, scopoletin, d-sedoheptulose, succinic acid, and xylitol in B. ceiba calyxes. The observed beneficial effect in our study might be attributed to the presence of these compounds in B. Ceiba calyxes. This is the first report presenting the antioxidant and antiglycation activities of B. ceiba calyxes and GC-MS analysis of active phytometabolites. These observations show that B. ceiba calyxes may become a potent and promising functional food to manage/control the development of diabetic complications.

Clinical validation of the chronic liver disease questionnaire for the Chinese population in Singapore.

BACKGROUND AND AIM: Assessment of health-related quality-of-life (HRQOL) in patients with chronic liver disease (CLD) requires the use of validated instruments that are understood by patients in their native language. We previously translated the Chronic Liver Disease Questionnaire into the Singapore-Mandarin version (CLDQ-SG). This study aims to examine the internal consistency and validity of the CLDQ-SG in patients with CLD. METHODS: We conducted a cross-sectional study of adult patients with CLD seen in a tertiary center in Singapore who completed both the CLDQ-SG and Short Form Health Survey 36 version 2 (SF-36v2) questionnaires. Internal consistency of the CLDQ-SG was assessed using Cronbach's alpha coefficient. Convergent and divergent validity of the SF-36v2 was assessed using the Spearman correlation coefficient, while discriminant validity was assessed using the Jonckheere-Terpstra test for trend. Exploratory factor analysis was performed to evaluate the factor structure of the CLDQ-SG. RESULTS: We enrolled 242 subjects (68.2% males, median age 67 years). Predominant etiology of CLD was chronic hepatitis B. Severity of CLD was divided into noncirrhotic (67.3%), compensated cirrhosis (24.0%), and decompensated cirrhosis (8.7%). Item convergent and discriminant validity of the CLDQ-SG was excellent, with 100% scaling success in all six domains. All domains exhibited good internal consistency, with Cronbach's α > 0.70. We observed a consistent trend of a reduction in mean CLDQ-SG score in the three groups reflecting the discriminant validity of the CLDQ-SG to assess changes in HRQOL in different severities of CLD. Factor analysis of the CLDQ-SG demonstrated an independent factor assessing sleep. CONCLUSION: The Singapore-Mandarin version of CLDQ-SG is a valid and reliable instrument to measure HRQOL in patients with CLD.

Understanding the metabolic perturbations in Carica papaya Linn. due to different ripening practices/agents using gas chromatography-mass spectrometry based metabolomics.

The study of fruit-ripening mechanism is vital as it plays a key role in the maintenance of fruit quality. Use of various xenobiotics for quick ripening has been shown to impact the quality of fruit, which in turn affect human health. In the present study, we made an attempt to understand the metabolic perturbations in Carica papaya Linn. (papaya), which has been ripened either by the ripening practice (room temperature process as control) and/or ripening agents (calcium carbide and ethylene) using gas chromatography-mass spectrometry (GC-MS) based metabolomics. The partial least squares-discriminant analysis has revealed significant alternations in 13 metabolites mainly sugars, amino acids, fatty acids, and organic acids as well as disturbances in five metabolic pathways due to different ripening practice/agents. The individual comparison of calcium carbide with control and ethylene with control has found 13 and 11 metabolites, respectively, which are common to the PLS-DA of three ripening groups. The GC-MS-based metabolomics has been able to predict the metabolic perturbations in papaya resulting from the ripening practice/agents. The findings from the present analysis has a wide application in food quality and will help to address safety concerns.

Evaluating the metabolic perturbations in Mangifera indica (mango) ripened with various ripening agents/practices through gas chromatography - mass spectrometry based metabolomics.

Mangifera indica L. (mango) is said to be the king of fruits due to its rich nutritional properties and mainly originates from the Indian sub-continent. The consumption pattern of the mangoes has increased drastically, due to which, many ripening practices/agents were used to make it ready-to-eat fruit or juice for the consumers. The fruit quality and metabolic composition are said to be altered due to different ripening agents/practices. The present communication mainly deals to understand the metabolic perturbations in mango fruits due to different ripening practices/agents (room temperature ripening, ethylene, and calcium carbide) using gas chromatography - mass spectrometry based metabolomics. The partial least square-discriminant analysis has found 16 differential metabolites for different ripening agents/practices which are belong to the classes of amino acids, fatty acids, sugars, and polyols. Four metabolic pathways were found to alter in the fruit metabolome due to different ripening agents/practices. Fructose, glucose, and galactose were found to be significantly up-regulated due to calcium carbide ripening in comparison to other ripening agents/practices. Overall findings from the present study advocates that mass spectrometry based metabolomics can be valuable tool to understand the fruit quality and safety with respect to consumer health.

A Rapid Method for the Quantitative Determination of 34 Pesticides in Nonalcoholic Carbonated Beverages Using Liquid-Liquid Extraction Coupled to Dispersive Solid-Phase Cleanup Followed by Gas Chromatography with Tandem Mass Spectrometry.

An economical, rapid, and sensitive multiresidue method using liquid-liquid extraction (LLE) coupled with dispersive SPE (dSPE) cleanup was developed for the quantitative determination of 34 multiclass multiresidue (MCMR) pesticides (14 organochlorines, eight organophosphates, 10 synthetic pyrethroids, and two herbicides) in nonalcoholic carbonated beverages (cola, orange, lemon-lime, and citra) using GC with tandem MS. The procedure mainly involved LLE by dichloromethane and dSPE cleanup in the presence of magnesium sulfate, primary secondary amine, and C18. The RSD of the developed method was found to be less than 14%. The LOD and LOQ values for all the analyzed pesticides were found in the ranges of 0.001-0.027 µg/L and 0.004-0.088 µg/L, respectively. The LOQ levels of the pesticides analyzed were found to be well below the recommended limit by the European Union (0.1 µg/L in water). The mean recoveries of pesticides in different nonalcoholic carbonated beverages (cola, orange, lemon-lime, and citra) were found to be in the range of 79-111%, with RSDs less than 11%. The validation data prove that the method can be acceptable to regulatory agencies for the routine analysis of MCMR pesticides in nonalcoholic carbonated beverages.

Picolinic acid based Cu(II) complexes with heterocyclic bases--crystal structure, DNA binding and cleavage studies.

In view of the importance of picolinic acid (PA) in preventing cell growth and arresting cell cycle, new PA based metallonucleases were designed with a view to study their DNA binding and cleavage abilities. Three new Cu(II) complexes [Cu(II)(DPPA)].4H2O (1),[Cu(II)(DPPA)(bpy)].5H2O (2) and [Cu(II)(DPPA)(phen)].5H2O (3), were synthesized using a picolinic acid based bifunctional ligand (DPPA) and heterocyclic bases (where DPPA: Pyridine-2-carboxylic acid {2-phenyl-1-[(pyridin-2-ylmethyl)-carbonyl]-ethyl}-amide; bpy: 2, 2'-bipyridine and phen: 1, 10-phenanthroline). DPPA was obtained by coupling 2-picolinic acid and 2-picolyl amine with l-phenylalanine through amide bond||. Complexes were structurally characterized by a single crystal X-ray crystallography. The molecular structure of 1 shows Cu(II) center essentially in a square planar coordination geometry, while complex 2 shows an approximate five coordinated square-pyramidal geometry. Eventhough we could not isolate single crystal for complex (3), its structure was established based on other techniques. The complex (3) also exhibits five coordinate square pyramidal geometry. The complexes show good binding affinity towards CT-DNA. The binding constants (Kb) decrease in the order 1.35 ± 0.01 × 10(5) (3) > 1.23 ± 0.01 × 10(5) (2) > 8.3 ± 0.01 × 10(4) (1) M(-1). They also exhibit efficient nuclease activity towards supercoiled pUC19 DNA both in the absence and presence of external agent (H2O2). The kinetic studies reveal that the hydrolytic cleavage reactions follow the pseudo first-order rate constant and the hydrolysis rates are in the range of (5.8-8.0) × 10(7) fold rate enhancement compared to non-catalyzed double stranded DNA (3.6 × 10(-8) h(-1)).